Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System

Protocol for Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 System

Summary: This protocol uses endonuclease-dead, programmable RNA-guided RNA-targeting Cas13 RNases (d)Cas13 proteins fused with fluorescent proteins to visualize and track RNA dynamics in live cells. This protocol details several aspects of the procedure, including gRNA design, fluorescent protein se...

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Bibliographic Details
Main Authors: Yang Wang, Liang-Zhong Yang, Ling-Ling Chen
Format: Article
Language:English
Published: Elsevier 2020-06-01
Series:STAR Protocols
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166720300241
Description
Summary:Summary: This protocol uses endonuclease-dead, programmable RNA-guided RNA-targeting Cas13 RNases (d)Cas13 proteins fused with fluorescent proteins to visualize and track RNA dynamics in live cells. This protocol details several aspects of the procedure, including gRNA design, fluorescent protein selection, nuclear localization signal adjustment, raw data analysis, operation steps, and extended optional applications that have been successfully applied in the visualization of NEAT1, SatIII, MUC4, and GCN4 RNAs.For complete information on the use and execution of this protocol, please refer to Yang et al. (2019).
ISSN:2666-1667

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